Modular parallel/serial dual microfluidic chip

ABSTRACT

A system for testing a treatment agent for a biologic material includes an input for receiving a biologic sample. A plurality of micro-pumps pump a portion of the biologic sample from the first reservoir into a connected module. A first module includes a first plurality of testing pathways for testing a first portion of the biologic sample. A first module connector removeably connects the first module to the distributor module. A second module includes a second plurality of testing pathways for testing a second portion of the biologic sample. The selected pathway applies at least one dosage level of a treatment agent to the second portion of the biologic sample. A second module connector removeably connects the second module to the distributor module, wherein treatment agent and the plurality of dosage levels tested by the system may be selected by selecting the second module associated with the second module connector.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority to and the benefit of U.S. ProvisionalApplication No. 62/584,655, filed Nov. 10, 2017, and entitled MODULARPARALLEL/SERIAL DUAL MICROFLUIDIC CHIP, the contents of which areincorporated by reference herein in their entirety.

TECHNICAL FIELD

The present invention pertains in general to a microfluidics lab-on-chipsystem and, more particularly, to the use of a microfluidics chip andtesting at the point of care.

BACKGROUND

The emergence and spread of antibiotic-resistant bacteria are aggravatedby incorrect prescription and use of antibiotics. Courts have thisproblem is the fact that there is no sufficiently fast diagnostic testto guide correct antibiotic prescription at the point of care.Currently, some fluid sample is retrieved from a patient and forwardedto a lab for testing to determine a specific treatment regimen. As asafeguard, the patient is sometimes initially given large doses of ageneral antibiotic until a more specific antibiotic can be determined totarget the specific bacteria. This can take upwards of two or threedays, as the process requires growing the bacteria in some culturemedium and observing its response to various antibiotics.

SUMMARY

The present invention disclosed and claimed herein, in one aspect,comprises a system for testing a treatment agent for a predeterminedbiologic material having a distributor module includes an input forreceiving a biologic sample containing a unique combination of apredetermined biologic material that must be treated via one of aplurality of treatment agents and a patient's biologic material. A firstreservoir holds the biologic sample containing the unique combination ofthe predetermined biologic material. A plurality of micro-pumps pumps aportion of the biologic sample from the first reservoir into a connectedmodule. A first module includes a first plurality of testing pathwaysfor testing a first portion of the biologic sample containing the uniquecombination of the predetermined biologic material and the patient'sbiologic material. Each of the plurality of first parallel testingpathways applies a different treatment agent of a plurality of treatmentagents to the first portion of the biologic sample. A first moduleconnector removeably connects the first module to the distributormodule, wherein the plurality of treatment agents tested by the systemmay be selected by selecting the first module associated with the firstmodule connector. A second module includes a second plurality of testingpathways for testing a second portion of the biologic sample containingthe unique combination of the predetermined biologic material and thepatient's biologic material in a selected pathway of the secondplurality of testing pathways. The selected pathway selected responsiveto a control input responsive to results of the first plurality oftesting pathways. The selected pathway applies at least one dosage levelof a treatment agent to the second portion of the biologic sample. Asecond module connector removeably connects the second module to thedistributor module, wherein treatment agent and the plurality of dosagelevels tested by the system may be selected by selecting the secondmodule associated with the second module connector.

BRIEF DESCRIPTION OF THE DRAWINGS

For a more complete understanding, reference is now made to thefollowing description taken in conjunction with the accompanyingDrawings in which:

FIG. 1 illustrates a high-level view of a microfluidics chip of thepresent disclosure;

FIGS. 2A-2C illustrate detailed views of the multiple stages of analysisprovided by the microfluidics chip of FIG. 1;

FIGS. 3A-3D illustrate diagrammatic views of the various cell captureregions and the interspersed pumps for the microfluidics chip of FIG. 1;

FIGS. 4A-4G illustrates detailed views of the first viewing stage;

FIG. 5 illustrates a detailed view of the first parallel driving stage;

FIGS. 5A and 5B illustrate details of the coating applied to the microchannels in the first driving stage;

FIG. 6 illustrates a detail of the serial driving stage;

FIGS. 7A-7D illustrate detailed views of a valveless nozzle/diffusermicropump;

FIG. 8 illustrates a detailed view of a piezoelectric micropump;

FIG. 9 illustrates a detailed view of a multi-chamber micropump withcheck valves;

FIG. 10 illustrates a flowchart for the high-level operation of themicrofluidics chip;

FIG. 11 illustrates a flowchart for the initial loading operation of thefluid sample;

FIG. 12 illustrates a flowchart for the viewing or cell counter stage ofanalysis;

FIGS. 13A-13C illustrate diagrammatic use for the cell counter;

FIG. 14 illustrates a flowchart for the main parallel stage of analysis;

FIG. 15 illustrates the serial stage of analysis;

FIG. 16 illustrates a simple fight diagrammatic view of themicrofluidics chip;

FIG. 17 illustrates a simplified diagrammatic view of a parallel module;

FIG. 18 illustrates simplified diagrammatic view of a serial module;

FIG. 19 illustrates a simplified diagrammatic view of a serial modulearranged in parallel;

FIGS. 20A and 20B illustrated a diagrammatic view of an embodimentutilizing a chemostat;

FIG. 21 illustrates a diagrammatic you have the microfluidics chiputilizing valves;

FIGS. 22A and 22B illustrate cross-sectional views of a micro valve

FIG. 23 illustrates a diagrammatic view of preparing a biologic sampleand disposing it in the well on the microfluidic chip;

FIG. 24 illustrates a cross-sectional view of an RT-lamp interfaced witha cell phone;

FIG. 25 illustrates a perspective view of the RT lamp interfaced with amicrofluidic chip and a cell phone;

FIG. 26 illustrates a side view of a cell phone interfacing with themicro fluidic chip;

FIG. 27 illustrates a window view of the camera and the alignmentprocess;

FIGS. 28A-28H illustrate multiple views of a diagram of the microfluidicchip in schematic form and various loading and analysis steps associatedthere with;

FIG. 29 illustrates a flowchart for the overall analysis processutilizing the microfluidic chip;

FIG. 30 illustrates a flowchart to pick in the details of the test path;

FIG. 31 illustrates a perspective view of a micro modular microfluidicchip;

FIG. 32 illustrates a cross-sectional view in detail of theinterconnection in the modular microfluidic chip;

FIG. 33 illustrates a top view of the system for receiving multiplemodules; and

FIG. 34 illustrates a top view of the micropump module showing thedistribution of microchannels.

DETAILED DESCRIPTION

Referring now to the drawings, wherein like reference numbers are usedherein to designate like elements throughout, the various views andembodiments of a Modular Parallel/Serial Dual Microfluidic Chip isillustrated and described, and other possible embodiments are described.The figures are not necessarily drawn to scale, and in some instancesthe drawings have been exaggerated and/or simplified in places forillustrative purposes only. One of ordinary skill in the art willappreciate the many possible applications and variations based on thefollowing examples of possible embodiments.

Referring now to FIG. 1, there is illustrated a diagrammatic view of amicrofluidics chip 102 at a high-level view. There is provided in themicrofluidics chip 102 an input stage 104 that is operable to receive abiological specimen. As used herein, a “sample” must be capable offlowing through microfluidic channels of the system embodimentsdescribed hereinbelow. Thus, any sample consisting of a fluidsuspension, or any sample that be put into the form of a fluidsuspension, that can be driven through microfluidic channels can be usedin the systems and methods described herein. For example, a sample canbe obtained from an animal, water source, food, soil, air, etc. If asolid sample is obtained, such as a tissue sample or soil sample, thesolid sample can be liquefied or solubilized prior to subsequentintroduction into the system. If a gas sample is obtained, it may beliquefied or solubilized as well. The sample may also include a liquidas the particle. For example, the sample may consist of bubbles of oilor other kinds of liquids as the particles suspended in an aqueoussolution.

Any number of samples can be introduced into the system for analysis andtesting, and should not be limited to those samples described herein. Asample can generally include any suspensions, liquids, and/or fluidshaving at least one type of particle, cellular, droplet, or otherwise,disposed therein. In some embodiments, a sample can be derived from ananimal such as a mammal. In a preferred embodiment, the mammal can be ahuman. Exemplary fluid samples derived from an animal can include, butare not limited to, whole blood, sweat, tears, ear flow, sputum, bonemarrow suspension, lymph, urine, brain fluid, cerebrospinal fluid,saliva, mucous, vaginal fluid, ascites, milk, secretions of therespiratory, intestinal and genitourinary tracts, and amniotic fluid. Inother embodiments, exemplary samples can include fluids that areintroduced into a human body and then removed again for analysis,including all forms of lavage such as antiseptic, bronchoalveolar,gastric, peritoneal, cervical, athroscopic, ductal, nasal, and earlavages. Exemplary particles can include any particles contained withinthe fluids noted herein and can be both rigid and deformable. Inparticular, particles can include, but are not limited to, cells, aliveor fixed, such as adult red blood cells, fetal red blood cells,trophoblasts, fetal fibroblasts, white blood cells, epithelial cells,tumor cells, cancer cells, hematopoeitic stem cells, bacterial cells,mammalian cells, protists, plant cells, neutrophils, T lymphocytes,CD4+, B lymphocytes, monocytes, eosinophils, natural killers, basophils,dendritic cells, circulating endothelial, antigen specific T-cells, andfungal cells; beads; viruses; organelles; droplets; liposomes;nanoparticles; and/or molecular complexes. In some embodiments, one ormore particles such as cells, may stick, group, or clump together withina sample.

In some embodiments, a fluid sample obtained from an animal is directlyapplied to the system described herein at the input stage, while inother embodiments, the sample is pretreated or processed prior to beingdelivered to a system. For example, a fluid drawn from an animal can betreated with one or more reagents prior to delivery to the system or itcan be collected into a container that is preloaded with such a reagent.Exemplary reagents can include, but are not limited to, a stabilizingreagent, a preservative, a fixant, a lysing reagent, a diluent, ananti-apoptotic reagent, an anti-coagulation reagent, an anti-thromboticreagent, magnetic or electric property regulating reagents, a sizealtering reagent, a buffering reagent, an osmolality regulating reagent,a pH regulating reagent, and/or a cross-linking agent.

At this point in the process, a finite amount of biofluids is disposedin the reservoir ready for transferring to subsequent stages. Thisamount of fluid is then transferred to another stage via a driving stage106 in order to transfer this biofluid to another reservoir, thatassociated with a viewing stage 108. At this stage, a technician canexamine the biofluid and determine the makeup of the biofluid,discriminate cells, etc. in order to make certain decisions as to goingforward with remaining tests. The microfluidic chip then transfers thebiofluid at the viewing stage 108 to a parallel analysis stage 115through a parallel driving stage 110 wherein the biofluid is dividedamong a plurality of parallel path this for analysis of the reaction ofthe material in the biofluid with different reagents in a reading. Thisrequires a certain amount of the biofluid to be transferred to thisanalysis stage. Thereafter, a decision is made as to whether to transferthe remaining biofluid from the viewing stage 108, in order to performmore testing and/or analysis on the biofluid. At this stage the process,only one of the multiple second stage or serial stage path is selected.One reason for this is that there is only a finite amount of biofluidavailable and there is no need for testing along paths that areassociated with previous decisions indicating that the results will benegative along these paths. Each of these serial passes associated withone of the parallel paths. Thus, if there are five parallel paths, therewill be five serial paths. Note that the term “serial path” is a termmeaning that it is within the serial decision tree and it need notactually be a plurality of serial paths that are linked together in aserial manner, although they could be and are in some embodimentsdescribed hereinbelow. It is necessary to perform the testing/analysisalong each of the five parallel paths, but a decision at this pointindicates that only one of the serial paths will be required for thetesting/analysis purpose. This will be described in more detailhereinbelow.

Referring now to FIGS. 2A-2C, there are illustrated diagrammatic viewsof the various stages of the process. With specific reference to FIG.2A, there is illustrated a diagrammatic view of first viewing stage,wherein the amount of biofluid stored in the input stage reservoir 104is driven to the viewing stage 108 reservoir. At this stage, opticaldevice 202, for example, can be used to view the cells disposed withinthe medium. This medium could actually be the actual biofluid that wasprovided in the sample from the human/animal or could be some dilutedversion thereof. However, this biofluid will contain some cellularmaterial or some particulate of interest. This can be viewed with theout device 202 and then passed to a processor 204, or a human couldanalyze the results. With utilization of the processor 204, the actualform of biofluid, and analog form, is transferred to a digital form.This could be in the form of cell counting for verification of aparticular cell. As will be described hereinbelow, affinity labels canbe associated with each of the cells or particulates in the biofluid andthis could facilitate visual recognition of different characteristics ordifferent types of cells, such as proteins, bacteria, etc. Each of thesecellular materials can have a particular affinity label associated therewith that allows it to be visually identified via some characteristicssuch as florescence or even magnetic properties associated with theaffinity label. Again, this will be described hereinbelow. Although anoptical device 202 was illustrated and described, any other type ofdevice for analyzing the characteristics of a particular affinitylabeled cell can be utilized, such as some type of magnetometer, etc.

Referring now to FIG. 2B, there is illustrated the next parallel drivestage. At this stage, a micropump is utilized in the parallel drivestage 110 to pump at least a portion of the biofluid stored in thereservoir associated with the viewing stage 108 is transferred to all ofthe parallel reading/analysis paths. In this step, it can be seen that aportion of the biofluid in the reservoir associated with the viewingstage 108 and is biofluid exists in each of these parallel paths foranalysis. There is an indication in one of these parallel paths,associated with the reservoir 210, that shows a positive indication of areaction of some type that is viewable. If, for example, this werebacteria, one reagent could be an antibiotic in a large dosage thatwould destroy the particular target bacteria and this would berecognized by an observer. The other three paths, associated withreservoirs 214, 216 and 218 (an example of 4 paths), would have noreaction and, as such, would not have affected the bacteria associatedtherewith. In this example, a high level of concentrated antibiotic isprovided that would destroy the bacteria, but at this level of analysis,there is no indication provided as to the actual dosage of thatantibiotic that would destroy the bacteria, other than the fact that alarge dosage of this particular antibiotic will destroy the targetbacteria. It is important to keep in mind that this particular biofluidmay have multiple and different bacteria, proteins, etc. containedtherein.

Referring now to FIG. 2C, there is illustrated a diagrammatic view ofthe final serial stage of analysis/testing. Since the first stage oftesting/analysis transferred some of the biofluid from the viewing stage108 to the parallel stages 114, there is still some biofluid remainingin the viewing stage 108. This is a selectively transferred to one ofthe serial paths, that associated with the testing reservoir 210. Thereare provided a plurality of bypass channels 220 associated with each ofthe serial paths and only the bypass channel 220 associated with thereservoir 210 in the parallel path 114 will be selected for transferringbiofluid to this particular serial path associated with the reservoir210 for testing. It will first be pumped to be a micropump in a serialdrive stage 222 to a first serial reservoir 224 for testing/analysis. Ifthe test is negative, it can then be passed to a subsequent serialdriving stage 226 to a subsequent serial reservoir 228 fortesting/analysis and so on. As will be described hereinbelow, there canbe provided a single bypass path 220 which is connected to a manifoldassociated with each of the serial paths and each of the manifolds canbe associated with each of the different reservoirs for testing, i.e.,at this point the testing is parallel to all of the subsequent testingreservoirs. In the mode illustrated in this FIG. 2C, it is necessary totransfer all of the necessary biofluid, i.e., typically the remainingbiofluid in the viewing stage reservoir 108, to the reservoir 224 andpass all of that biofluid to the next reservoir 228 and so on. Thus, ateach stage, all of the biofluid transferred in the subsequent stages istested at each subsequent stage. In a parallel configuration, theremaining biofluid in the viewing stage 108 would be required to bedivided among the different testing reservoirs at each of the subsequentstages. This will be described in more detail hereinbelow.

Referring now to FIGS. 3A-3D, there are illustrated diagrammatic viewsof the process and fluid flow. In FIG. 3A come there is illustrated anoverall process flow for the embodiment described hereinabove. Thisembodiment, there is provided an input well 302 for receiving thebiologic sample indicated by numeral 303. This constitutes a finitevolume that must be transferred via a micropump to a viewing reservoir306. At this point, substantially all of the biofluid is transferredfrom the reservoir 302 to the viewing reservoir 306. This is the firststage of the process. The second stage of the process is illustrated asproviding three separate testing reservoirs 308, 310, 312, attached atone to a microchannel manifold 314. Each of the testing reservoirs 308,310, 312, as will be described hereinbelow, is comprised of a serpentinemicrochannel 316 attached at one end to the manifold 314 and at theother end to a viewing reservoir 318. A micropump 320 is provided fortransferring biofluid from the viewing reservoir 306 to the manifold314. This will be divided among the three testing reservoirs 308, 310,312 and substantially even amounts. The biofluid will traverse theserpentine microchannel 316, which is coated with a particular reagent,one example being an antibiotic. In this example, the antibiotic is at avery high concentrated level, each of the testing reservoirs 308, 310and 312 having a different antibiotic associated there with. Only aportion of the biofluid in the viewing reservoir 306 will be transferredto these three testing reservoirs 308, 310 and 312 for testing/analysisand viewing at the associated viewing reservoir 318. The serpentineshape, when used with a medium containing cells such as in a biologicsample, facilitates and enhances mixing due to the increased interfacialcontact area between the cells within the biofluid sample.

The next step of testing/analysis will be selected only upon a positivetest occurring within one of the three testing reservoirs 308, 310 and312. However, each of the testing reservoirs 308, 310 and 312 hasassociated there with a subsequent group of testing reservoirs. In thisembodiment, each of the subsequent testing reservoirs is comprised of aplurality of sub reservoirs 330, each of the sub reservoirs 330 beingconfigured identical to the testing reservoirs 308, 310 and 312, with aserpentine microchannel region 316 and a viewing reservoir 318. A singlebypass microchannel 220 is provided to connect viewing reservoir 306 toa sub reservoir manifold 332. Each of the particular sub reservoir pathshave associated there with a separate micropump 334. Only one of thesemicropumps 334 is selected for transferring the remaining portion of thebiofluid stored in the viewing reservoir 306 to the selected path. Inthis embodiment, the remaining portion of the biofluid is transferred tothe first reservoir 330 bypassing the biofluid through the serpentinemicrochannel 316 to the associated viewing reservoir 318. Thisparticular microchannel will have coating of antibiotic, in this exampleabove, at a relatively low dose. If the bacteria, for example, do notreact accordingly with this level of antibiotic, it can be recognized assuch in the viewing reservoir 318. It is noted that the antibioticassociated with the coating on the walls of the microchannel 330 at thisdosage will not be picked up by the bacteria and, as such, the bacteriain the viewing reservoir 318 for the first sub reservoir 330 in theselected path will still be intact. It can then be pumped from thereservoir 318 associated with the first testing reservoir 330 in thechain to a subsequent testing reservoir 330 with a subsequent micropump336. This subsequent sub reservoir will have a concentration ofantibiotic in its serpentine microchannel 316 that is at a higher level.As the level increases, a gradient is tested for, such that the dosagecan be gradually increased until the bacteria are destroyed. If, forexample, the bacteria were associated with an affinity label that madeit fluoresce, this would be recognized. It could also be that there aremultiple bacterial types contained within the biofluid that are eachassociated with a different affinity label and this could be recognized.It could, in fact, the case that one type of bacteria perfected at afirst dosage level of the antibiotic and a second bacteria were affectedat a another dosage level of the antibiotic.

Referring now to FIG. 3B, there is illustrated a diagrammatic view of analternate process flow. This will work substantially identical to theembodiment of FIG. 3A, come up until the operation at the manifold 332associated with the sub reservoirs. In this embodiment, the threemicropumps 334 each feed a sub reservoir manifold 340. Each of the subreservoir manifolds 340 is connected to a plurality of the subreservoirs 330 associated with each path. In this embodiment, there areonly illustrated three sub reservoirs 330 for each of the sub reservoirmanifolds 340, although each path could have a different number of subreservoirs 330 associated therewith. The difference between these twoembodiments is that, at this point, the amount of biofluid remaining inthe viewing reservoir 306 now must be divided amongst all of the subreservoirs attached on one end thereof to the associated sub reservoirmanifold 340 selected by the activated one of the micropumps 334. Thiswill result in potentially less biofluid being available for thetesting/analysis step. This will also mean that each of the viewingreservoirs 318 associated there with will have a smaller volumeassociated therewith.

Referring now to FIG. 3C, there is illustrated a diagrammatic view thatprovides a simplified diagram of the transfer from reservoir toreservoir. In this illustration, the input stage is illustrated as aninput reservoir 350 labeled R0. A micropump 352 is operable to transferthe contents of this input reservoir, the biofluid, to a secondreservoir, a viewing reservoir 354, labeled R1. A portion of thecontents of this reservoir are then transferred via a micropump 356 to aplurality of parallel stage reservoirs 358 labeled R2. This is the firsttesting/analysis stage. After this stage, the remaining contents of theviewing reservoir 354 are transferred to the subsequent serial stagereservoirs via a pump 360 via a bypass path and microchannel 362. Theserial stage reservoirs are labeled R3, R4, etc. This illustration setsforth how the entire contents of the input reservoir R0 are transferreddown the chain. This is best illustrated in FIG. 3D. In thisillustration, it can be seen that entire contents of reservoir R0 aretransferred to reservoir R1. At this point, only a portion of thecontents are transferred to reservoir R2. The remaining contents aresequentially transferred to R3, R4, and so on. For this illustration,the entire remaining contents of the reservoir 354, R1, will betransferred down the chain entirely to reservoir R3, then to reservoirR4, and so on. In the alternate embodiment, as described hereinabove,and not illustrated in FIG. 3D, the bypass 362 could be connected toeach of the reservoirs R3, R4, etc. in parallel, noting that theremaining contents of the reservoir R1 will then be divided amongst theparallel connected reservoirs R3, R4, etc.

Referring now to FIGS. 4A-4G, there are illustrated diagrammatic viewsof the initial processing section associated with the viewing stage 108.There is provided a substrate 402 upon the surface of which are formed aplurality of wells and microchannels. A first well 404 is provided forreceiving the biofluid sample in this well has a finite volumeassociated there with. At the bottom of this well a microchannel 406extends outward and up to the surface to an opening 408. The purpose ofthis microchannel 406 extending to the bottom of the well 404 is toensure that the biofluid can be completely pumped from the well 404. Forthe formation of this microchannel 406, it might be that themicrochannel is formed through the surface of the substrate 402 and thena cover plate (not shown) having a surface that extends down into theopen microchannel. An adjacent channel 410 is disposed proximate theopening 408 to provide another opening therefore in order to accommodatea micropump 412 (shown in phantom) interface with the opening 408 andthe one end of the microchannel 410 for transferring fluid from the well404 to the microchannel 410. The microchannel 410 extends along thesurface of substrate 402 in order to interface with a viewingwell/reservoir 412. As the biofluid passes through the microchannel 410and the viewing well 412, a desired analysis can be performed on thecontents of the biofluid. As described hereinabove, in one example,various cells in the biofluid might consist of different types ofbacteria, proteins, etc. and each of these may have associated therewith a specific affinity label, which is optically detectable. Thereare, of course, other means by which affinity labels can be detected. Asthe cells contained within the biofluid pass through the viewingwell/reservoir 414, they can be examined. The viewing well/reservoir 414on the other side thereof is connected to one side of a microchannel416, the other side thereof connected to a reservoir 418. Since themicropump 412 must force the biofluid through the microchannels and theviewing well/reservoir 414, there is required the necessity for aholding reservoir 418 to be present. However, initially, this reservoir418, the microchannel 410 and the viewing well/reservoir 414 will haveair disposed therein. This air must be removed. This can be done with anegative pressure of some sort or just a waste gate output to theatmosphere. This is provided by a waste gate microchannel 420 that isconnected to an opening 422 through the cover glass (not shown) or tothe side of substrate 402. A valve 423 could be provided above theopening 422. As biofluid enters the reservoir 418, air will be pushedout through the microchannel 420. It is desirable for this microchannel422 to have as low a profile as necessary such that only air exitstherefrom. Depending upon the size of the cells contained within thebiofluid, the microchannel 420 can be significantly smaller and have alower profile than the microchannels 410 and 416. Is important to notethat, once the micropump 412 transfers the biofluid from the well 404,the volume transferred will be spread between the two microchannels 410and 416, the viewing well 414 and the reservoir 418. Thus, the reservoir418 has a significantly larger volume that any of the microchannels 410and 416 and the viewing well/reservoir 414. Additionally, it may be thatthe depth of the wells/reservoirs 404 and 418, as well as the viewingwell reservoir 414 are also as shallow as the microchannels 410 and 416but significantly wider to accommodate the required volume.

The outlet of the reservoir 418 is connected from the bottom thereofthrough a microchannel 426 to an opening 428 on the upper surface of thesubstrate 402. This is interfaced with a micropump 430 (in phantom) toan adjacent microchannel 432 for subsequent processing. These micropumps412 and 430, although illustrated as being flush with the substrate,will typically be disposed above the cover plate (not shown) with holesdisposed through the cover plate. The opening 428 will be a horizontalmicrochannel associated with the manifold 314 described hereinabove.This will be associated with a plurality of micropumps 430 for each ofthe parallel paths or the bypass path. A cross-sectional view of theembodiment of FIG. 4A is illustrated in FIG. 4B, with a cover plate 440disposed over the substrate 402 with an opening 442 disposed above thewell 404 for receiving the biofluid sample.

FIGS. 4C and 4D illustrate top view and cross-sectional views of thereservoir 418 illustrating how the microchannel 416 feeds biofluid intothe top of the reservoir 418, and the flow path for the biofluid fromthe reservoir 418 through the microchannel 426 from the bottom of thereservoir 418. However, it may be that, with capillary action, the depthof the reservoir 418 could be equal to that of the microchannels 416 and426 such that they are all at the surface of the substrate 402 for easeof manufacturing. When a negative pressure is placed upon the reservoir418, air will be pulled into the microchannel 426 through themicrochannel 420. It is possible in this mode that the micropump 412could be operated to actually create a positive pressure in themicrochannel 416 to force the biofluid in the reservoir 418 into theopening 428 through the microchannel 426. Again, the microchannel 420would preferably have a dimension that was smaller than the smallestcell size within the biofluid.

Referring now to FIGS. 4E and 4F, there are illustrated top view andcross-sectional views of the reservoir 418 with an alternate embodimentillustrating microchannel 426′ as being beneath the bottom of thereservoir 418 to allow more complete emptying of the reservoir 418.

Referring now to FIG. 4G, there is illustrated an alternate embodimentof inlet wells for receiving the biofluid sample. There is provided thewell 404 for receiving the biofluid sample and a second well 464receiving an additional fluid sample. This fluid sample in well 460could be some type of dilutant or it could be a medium containingvarious affinity labels. As noted hereinabove, the fluid sample couldhave associated there with affinity labels prior to the biofluid samplebeing disposed in the well 404. However, it is possible that themicrofluidic chip have disposed in the well 460 a medium containingaffinity labels, for example. The well 460 would be interfaced through amicrochannel 462 to an opening 464 adjacent the opening 408. A twoinput, one output, micropump 412′ that interfaces with the microchannel410.

Referring now to FIG. 5, there is illustrated a diagrammatic view of themicrochannel structure associated with the parallel stage of operation.The microchannel 426 is interfaced with a microchannel manifold 502which corresponds to the opening 428. This microchannel manifold 502 isinterfaced with a plurality of micropumps 504, corresponding to themicropump 430. These micropumps 504 are disposed in pairs, each pairassociated with one testing reagent. As noted hereinabove, there areprovided a plurality of parallel paths, each associated with a reservoir312 having a serpentine microchannel 316 and a viewing reservoir 318.The first micropump 504 in the pair of micropumps 504 is connected toone end of the associated serpentine microchannel 316. When thismicropump 504 is activated, biofluid from the reservoir 418 is passedthrough the manifold microchannel 502 and through the serpentinemicrochannel 316 to the viewing reservoir 318. As was the case above,there is provided a waste microchannel 506 for each of the reservoirs318 to allow air to escape therefrom as biofluid is forced through themicrochannel 316. The micropump 504 associated with this serpentinemicrochannel 316 will be operated for a sufficient amount of time totransfer sufficient biofluid from the reservoir 418 through theserpentine a channel 316 and finally into the reservoir 318 to fill thereservoir 318. The microchannel 506 can have some type of valveassociated with the opening thereof to prevent the escape of anybiofluid therefrom or, alternatively, the dimensions of thatmicrochannel 506 could be small enough to prevent any appreciable amountof cells escaping therefrom. Although not illustrated, the one of thepair of micropumps 504 associated with the parallel stage of operationand associated reservoirs 312 will also be operated to fill theassociated serpentine microchannel 316 and reservoir 318.

Referring now to FIGS. 5A and 5B, there are illustrated cross-sectionalviews of the serpentine microchannel 316. As described hereinabove, thesides of these channels 316 are coated with some type of reagent. Forexample, if a Urinary Tract Infection (UTI) were suspected and werebeing tested for in the microfluidic chip, the sensitivity for commonantimicrobial agents for UTI treatment might include ampicillin (AMP),ciprofloxacin (CIP), and trimethoprim/sulfamethoxazole (SXT), thesebeing three agents that could be tested for and three different paths.The bacteria that might exist within the urine samples from anindividual could be any of uropathogenic E. coli strains (EC132, EC136,EC137, and EC462). Some prior research has shown that, throughantimicrobial resistance profiles of these pathogens that EC132 isresistant to AMP and CIP but not SXT. EC136 is resistant to AMP only.EC137 is sensitive to all the antibiotics tested. EC462 is resistant toAMP and SXT but not CIP. In order to coat sides of the serpentinemicrochannels 316, one technique would to have a certain amount of theantibiotic dissolved in sterile water to the serpentine microchannels316 at different levels. Subsequently, the diluted antibiotic is driedby incubation at a desired temperature and desired time. The originaldiluted antibiotic has a starting concentration of a predetermined μg/mlconcentration. The surface area is sufficiently covered such that, whenthe biofluid passes thereover, it will interact with reagent.

Referring now to FIG. 6, there is illustrated a microchannel diagram ofthe reservoir 330 on the surface of the chip 402. This is connected bythe microchannel 507 from the associated one of the micropumps 504.After the results in the viewing reservoir 318 have been determined toyield a positive result, for that particular path in the parallelanalysis/testing operation, the other of the pair of micropumps 504 isactivated and the remaining amount of micro-fluid from the reservoir 418is transferred to the reservoir 330. This will be passed through theserpentine microchannel 316 and stored in the reservoir 318, labeled 602in FIG. 6. This is substantially larger than the reservoir 318associated with the reservoir 312. Thus, for this embodiment, theremaining portion of the biofluid from the reservoir 418 will besubstantially stored in the reservoir 602. This will have associatedthere with a waste microchannel 604 and an outlet microchannel 608 thatextends outward from the bottom of the reservoir 602 and up to anopening 610 in the surface of the substrate for interface with themicropump 336. The micropump 336 is operable, at the next stage of thetesting/analysis, to move the contents of the reservoir 602 over to thenext reservoir 330 for testing at that next concentration levelassociated with the next reservoir 330 in the sequence.

Referring now to FIGS. 7A-7D, there is illustrated an example of avalveless MEMS micropump. The micropump includes a body 702 with twopumping chambers 704 and 706. At the inlet side of each of the chamber704 and 706 is disposed a conical inlet 710 and 712, respectively. Theconical inlets 710 and 712 are wider at the pump chamber side andnarrower at the inlet side thereof. The inlet sides of conical inlet 710and 712 are connected to respective inlet channel 714 and 716. Theoutlet side of the chambers 704 and 706 are interfaced with conicaloutlets 718 and 720, respectively, the conical outlets 718 and 720having a narrower portion at the outlet of the respective pump chamber704 and 706 and a wider portion at the respective outlet thereofinterfacing with respective outlet channels 722 and 724. The conicalinlets 710 and 712 and outlets 718 and 720 are frustro conical in shape.A piezoelectric membrane and actuator 726 is dispose between the twopumping chambers 704 and 706 and is operable to be extended up into oneof the chambers 704 and 706 at one time to increase the pressure thereinand at the same time extend away from the other of the chambers 704 and706 to decrease the pressure therein. The operation is then reversed.

The piezoelectric membrane and actuator 726 is comprised of apiezoelectric disc 740 on one side of a membrane 742 and a piezoelectricdisc 744 on the other side thereof. Each of the piezoelectric discs 740and 744 are formed by stratifying a layer of use electric material 748between two layers of conducting material 750. Piezoelectric material748 can be made with Piezo Material Lead Zirconate Titanate (PZT-SA),although other piezoelectric materials can be used. The conductingmaterial 60 may be composed of an epoxy such as commercially availableEPO-TEK H31 epoxy. The epoxy serves as a glue and a conductor totransmit power to the piezoelectric discs 750. The piezoelectric discs750 are secured to the surface of the intermediate layer 748, so thatwhen a voltage is applied to the membrane 742, a moment is formed tocause the membrane 742 to deform.

The operation of the micropump will be described with reference to FIG.7D. At rest, the upper chamber 704 and the lower chamber 706 areseparated by a diaphragm pump membrane 742. The diffuser elements 710,712, 718 and 720 are in fluid communication with each respectivechamber. Diffuser elements are oriented so that the largercross-sectional area end of one diffuser element is opposite the smallercross-sectional area end of the diffuser element on the other side ofthe chamber. This permits a net pumping action across the chamber whenthe membrane is deformed.

The piezoelectric discs are attached to both the bottom and the top ofthe membrane. Piezoelectric deformation of the plates is varied byvarying the applied voltage so as to excite the membrane with differentfrequency modes. Piezoelectric deformation of the cooperating platesputs the membrane into motion. Adjustments are made to the appliedvoltage and, if necessary, the choice of piezoelectric material, so asto optimize the rate of membrane actuation as well as the flow rate.Application of an electrical voltage induces a mechanical stress withinthe piezoelectric material in the pump membrane 742 in a known manner.The deformation of the pump membrane 742 changes the internal volume ofupper chamber 704 and lower chamber 706. As the volume of the upperchamber 704 decreases, pressure increases in the upper chamber 706relative to the rest state. During this contraction mode, theoverpressure in the chamber causes fluid to flow out the upper chamber704 through diffuser elements on both sides of the chamber. However,owing to the geometry of the tapered diffuser elements, specifically thesmaller cross-sectional area in the chamber end of the left diffuserelement relative to the larger cross-sectional area of the rightdiffuser element, fluid flow out of the left diffuser element is greaterthan the fluid flow out the right diffuser element. This disparityresults in a net pumping of fluid flowing out of the chamber to theleft.

At the same time, the volume of the lower chamber 706 increases with thedeformation of the pump member 742, resulting in an under pressure inthe lower chamber 706 relative to the rest state. During this expansionmode, fluid enters the lower chamber 706 from both the left and theright diffuser elements. Again owing to the relative cross-sectionalgeometry of the tapered diffuser elements, fluid flow into the lowerchamber 706 through the right diffuser element is greater than the fluiddrawn into the lower chamber 706 through the left diffuser element. Thisresults in a net fluid flow through the right diffuser element into thechamber, priming the chamber for the pump cycle.

Deflection of the membrane 742 in the opposite direction produces theopposite response for each chamber. The volume of the upper chamber 704is increased. Now in expansion mode, fluid flows into the chamber fromboth the left and right sides, but the fluid flow from the rightdiffuser element is greater than the fluid flow from the left diffuserelement. This results in a net intake of fluid from the right diffuserelement, priming the upper chamber 704 for the pump cycle. Conversely,the lower chamber 706 is now in contraction mode, expelling a greaterfluid flow from the lower chamber 706 through the left diffuser elementthan the right diffuser element. The result is a net fluid flow out ofthe lower chamber 706 to the left.

Referring now to FIG. 8, there is illustrated a cross-sectional view ofa piezoelectric micropump with check valves. Membrane 802 is disposedwithin a pump chamber 804 and secured to a body 806. A piezoelectricdisc 808 is disposed beneath the membrane 802 and electrode 810 isdisposed below the piezoelectric disc 808. Deformation of the membrane802 with the piezoelectric disc at the appropriate frequency will causea volume of the pumping chamber 804 to change. An inlet valve 810 allowsfluid to flow into the chamber 804 and an outlet valve 812 allows fluidto flow out of the chamber 804.

Referring now to FIG. 9, there is illustrated a micropump 960 in which ananofabricated or microfabricated fluid flow pathway is formed betweenstructures. A first reservoir 961 terminates with a first gate valve 966which permits or restricts fluid flow between the first reservoir 961and a second reservoir 973. An electrolytic pump 985 drives a firstdiaphragm 965 which is communication with the second reservoir 973, toclose the first gate valve 966, and pulls a second diaphragm 969, whichopens a second gate valve 968 to drive fluid from the second reservoir973 to a third reservoir 973. The electrolytic pump 985 is driven byelectrowetting of a first membrane 962 on the first gate valve 916 sideof the pump. By switching to electrowetting of a second membrane 963, asdepicted in FIG. 16B, fluid within the third reservoir 973 is emittedfrom an exit opening 170 by actuation of the second diaphragm 969.

Referring now to FIG. 10, there is illustrated a flowchart depicting theoverall operation of the system. The process is initiated at a Startblock 1002 and then proceeds to a block 1004, wherein the biofluidsample is loaded. The process enclosed a block 1006, wherein thebiofluid is transferred to the viewing window or the cell counter. Theprocess then flows to a decision block 1008 to determine when thecounting operation is done, i.e., when the cells have beendiscriminated. As noted hereinabove, each of these cells could beassociated with, depending on upon the type, a particular affinity labelto allow them to be discriminated between within the viewing window. Theprocess then flows to a block 1010 in order to pump the biofluidmaterial to the next phase, that associated with the paralleltesting/analysis step. A decision is then made at a block 1012 as towhether this is a positive state, i.e., has any of the biofluid materialinteracted with a particular reagent to give a positive result. If not,the process is terminated at a block 1014 and, if so, the process flowsto a block 1016 in order to capture the biofluid material in a secondaryreservoir. Once the path is selected, the appropriate micropump isactivated and the biofluid material is pumped to the next reservoiralong the secondary path, as indicated by a block 1018. The process thenflows to a block 1022 in order to analyze the results at each secondaryreservoir and, if there is a positive result, as indicated by block1022, the process is terminated at a block 1024. If the result is notpositive, the process then flows to a block 1026 to determine if that isthe last testing reservoir and, if so, the process flows to theterminate block 1024. If there are more testing/analysis blocks throughwhich to process the biofluid material, the process then flows back tothe input of a block 1018 to pump the biofluid serial to the nexttesting reservoir.

Referring now to FIG. 11, there is illustrated a flowchart for theloading operation, which is initiated at a block 1101 and then flows toa block 1102 wherein the sample is placed in the well and then to adecision block 1104 to determine if this is a process wherein thebiofluid sample is to be mixed with some other diluted product or anaffinity label. If it is to be mixed, the process flows to a block 1106to mix the biofluid sample and, if not, the process bypasses this step.The process then flows to a block 1108 in order to activate the pump andtransferred the biofluid material after mixing to the next reservoir inthe process.

Referring now to FIG. 12, there is illustrated a flowchart for theprocess of the cell counting operation, i.e., the operation at theviewing reservoir. This is initiated at a block 1202 proceeds to a block1204 in order to transfer the biofluid material to the viewing chamber.The process enclosed a block 1206 in order to view the cells in realtime as they pass through the various microchannels and viewing window.The process then flows to a block 1208 in order to count the cells. Atthis stage, the cells can have various affinity labels associated therewith such that the target cells can be viewed and discriminated betweenbased upon the affinity labels associated therewith. If, for example,there were multiple types of bacteria contained within the biofluidsample and each of these types of bacteria had associated therewithdifferent affinity label that clips arrest at a different color, theykilled be discriminated between. Additionally, proteins would have adifferent affinity label than a bacteria and this would also allowdiscrimination between the two types of cells. The process then flows toa block 1210 to store the transferred biofluid in the reservoir and intoa block 1212 to terminate.

Referring now to FIGS. 13A-13C from their illustrated variousconfigurations for the cell counting operation. In the first embodimentof FIG. 13A, there are provided a three-part microchannel 1302, a middlesection microchannel 1304 and an outlet microchannel section 1306 themiddle section 1304 has a diameter that is slightly larger than thelargest cell that could be contained within the biofluid. This allowsthe cells to be transferred in a more orderly manner. The cell viewingwould be performed at this middle section microchannel 1304. In theembodiment of FIG. 13B, there are provided three varying diameter middlemicrochannel sections 1308, 1310 and 1312, each with different diametersto allow different size cells to flow therethrough. This type ofembodiment may facilitate some selection in the cells for viewing. Inthe embodiment of FIG. 13C, there is illustrated the above discloseembodiment wherein the microchannel 416 empties into the reservoir 418and the viewing is basically performed upon the cells within thereservoir 418.

Referring now to FIG. 14 come there is illustrated a flowchart for theparallel cell capture in the first testing/analysis stage. This isinitiated at a block 1402 and a process and proceeds to a block 1406 inorder to preload all of the cell capture areas having reagent associatedthere with, such that the portion of the biofluid stored in thereservoir 418 is transferred to the reservoirs associated with theparallel cell capture areas. The process enclosed a block 1408 whereinthe pump is activated to fill all of the cell capture wells associatedwith this stage of testing/analysis. The process then flows to a block1410 to possibly allow the cells to slowly go through the microchannelsin order to interact with the reagent. If so, this requires a certainamount of time and this would result in the micropumps operating at alower rate to allow sufficient time for the cells to flow through theserpentine microchannels 316 to interface with the particular coating onthe surfaces thereof. This basically is the amount of time required forthe micropumps to fill up the reservoir 318 associated there with. Thelength of the serpentine microchannel 316 would determine the amount oftime required to fill up the reservoir 318. Once the reservoir has beenfilled, as indicated by a block 1412, then the viewing window in thereservoir 318 is analyzed, as indicated by a block 1414. The path fromthe block 1410 to the input of the block 1414 indicates a path by whichthe micropumps can be run at a higher rate. The process then isterminated at a block 1416.

Referring now to FIG. 15, there is illustrated flowchart for the secondphase of the analysis, provided that the first phase indicated apositive result for one of the cell capture areas and the associatedreagent. This is initiated a block 1502 and then proceeds to a block1504 to preload all of the secondary cell capture areas with reagent andinto a function block 1506 to pump all of the remaining biofluidmaterial from the reservoir 418 into the first reservoir in thesecondary reservoirs 330. This also goes through and incubate step toallow the micropumps to pump at a slower rate to allow the biofluidmaterial to go through the serpentine microchannel 316 at a slower ratebefore it enters the associated reservoir 318. When the reservoir 318 isfilled, as indicate a by block 1510, the contents of the reservoir 318are analyzed at a block 1512. If the pump can be run at a faster rate,this is provided by a path around the block 1510. If the result ispositive, as indicated by a block 1514, then the process is terminatedat a block 1516. If not, the process flows from the block 1514 to ablock 1518 in order to the next reservoir 330 in the back to the inputof the serpentine microchannel 316 and then float the input of the block1508.

Referring now to FIG. 16, there is illustrated a simplified diagrammaticview of the microfluidics chip for processing a plurality of modules.The sample 303 is input to the well 302 and then pumped into the viewingwindow 306. A waste microchannel 1602 is provided an interface to theviewing window 306 that is interfaced with a micro valve 1604 to allowair to escape, or any bubbles that may be present, from the viewingwindow 306. Additionally, the waste microchannel 1602 could interfacewith an external vacuum source aid in fluid flow. A cellcounter/discriminator 1606 is provided for optically viewing thecontents of the viewing window 306, the output thereof processed via aprocessor 1608. The outlet of the viewing window 306 is interfaced witha manifold microchannel 1610 through a connecting channel 1612. At thispoint, the micro valve 1604 is closed such that the biofluid containedwithin the viewing window 306 and the interfaced with microchannelmanifold 1610 to allow fluid to be pump therefrom to a plurality ofdistribution paths along distribution microchannels 1614. It may be thatpump 304 would need to be activated in order to reduce the pressure atthe upper end of the viewing channel 306 or, alternately, a microchannel1618 interfaced with a micro valve 1620 could be provided to, when open,relieve the pressure in the upper end of the viewing window 306 to allowbiofluid to be pumped therefrom to the microchannel manifold 1610.

Each of the distribution microchannels 1614 is interfaced with aseparate module via an associated distribution pump 1624 to interfacewith and associated one of modules 1625, labeled A-Z, for example. Therecan be any number of modules provided. However, each module 1625 hasassociated there with a finite capacity and, therefore, the number ofmodules 1625 that can be interfaced to the viewing window 306 is afunction of the volume of biofluid contained therein and the capacity ofthe reservoirs of each of the individual modules 1625, each of theindividual modules 1625 potentially having a different capacity,depending upon the configuration thereof. However, selecting among thevarious distribution pump 1624 can allow desired tests to be done withthe available biofluid contained within the viewing window 306.

Referring now to FIG. 17 there is illustrated a diagrammatic view of oneof the modules 1625 associated with the parallel testing configuration,wherein biofluid is loaded into a plurality of testing reservoirs. Thedistribution pump 1624 associated there with transfers fluid from thedistribution microchannels 1614 to an intermediate microchannel manifold1702 which is then interface with a plurality of testing reservoirs 312,as described hereinabove. Each of these testing reservoirs has aserpentine microchannel 312 and a viewing window 318 associated therewith. As described hereinabove, each of these testing reservoirs canhave a different volume and a different configuration mechanically andcan be associated with a different test. They can each have a particularcoating of reagent, such as an antibiotic, to interact with the biofluidfor testing purposes to determine if there is any reaction of thebiofluid in the cells contained therein to the material coated on thesides of the serpentine microchannels 316. In the operation of thisparticular module 1625, all of these testing reservoirs 312 areassociated with different reagents and will be loaded in parallel. Forthis embodiment, will be desirable for each of the reservoir 312 to havethe same volume. If, however, they had different volumetric capacities,it would be necessary to have some type of waste gate with a micro valveto allow all of the viewing windows 318 to achieve full capacity.

Referring now to FIG. 18, there is illustrated a diagrammatic view ofthe serial wherein a plurality of testing reservoirs 330 is arranged ina series configuration. In this configuration, the associateddistribution pump 1625 will transfer biofluid from the microchannelmanifold 1610 through the distribution microchannels 1614 to the firstof the testing reservoirs 330. The biofluid will be contained within theviewing chamber 318 and, as noted hereinabove, there will be possible hesome type of waste microchannel associated micro valve to allowair/bubbles to escape during filling of the viewing window 318.Thereafter, a second serial pump 1706 is activated to transfer thecontents of the viewing window 318 to a second testing reservoir 330 inthe associated serpentine microchannel 316 and viewing window threeeight teen. In this transfer, there may be required a reliefmicrochannel (not shown) at the inlet end thereof to reduce the pressuretherein during the pumping operation. This will continue until all ofthe tests have been done. Each of the serpentine microchannels 316associated with each of the testing reservoirs 330 will have a graduatedincrease in the particular reagent to determine the dosage, in thisexample. It may be that, upon being exposed to the dosage of the reagentin the first testing reservoir 330 that cellular material in thebiofluid is somewhat affected by the reagent, i.e., the antibiotic, forexample. By moving to a higher concentration of the reagent in the nextsequential testing reservoir 330, this could be accounted for in theoverall analysis. It may be that the actual concentration in the nextsequential testing chamber 330 is not an exact incremental increase inthe reagent. For example, if it was desired to expose the biofluid toreagent increments of 10%, 20%, 30%, etc. in 10% increments, it may bethat the first testing chamber 330 has a concentration of 10% and thenthe second testing chamber has a concentration of possibly 16%,accounting for the fact that the accumulated effect of passing throughthe 10% testing chamber 330 and the 16% testing chamber 330 effectivelyprovides a 20% accumulated exposure in the second testing chamber 330and so on.

Referring now to FIG. 19, there is illustrated a diagrammatic view of aconfiguration for providing parallel loading of the serial configurationfor the incremental testing. This is similar to the embodiment of FIG.17, except that the testing chambers 330 are all interfaced with theassociated distribution pump 1625 through a microchannel manifold 1902in a parallel configuration, such that they are all loaded at the sametime, with each having a different concentration of reagent associatedthere with. In this configuration, however, since all of the testingchambers 330 will be loaded in parallel, there are required to be asufficient volume of biofluid contained within the viewing window 306initially to facilitate complete filling of each of the associatedviewing windows 318.

Referring now to FIGS. 20A-20B come there is illustrated a diagrammaticview of chemostat, wherein the associated distribution pump 1625transfers biofluid from the distribution microchannel 1614 two eightchemostat 2002. The details of the chemostat 2002 are illustrated inFIG. 20B. A main microchannel 2004 is interfaced on one and thereof withthe output of the distribution pump 1625 associated there with, with theother end of the microchannel 2004 interfaced with a waste gate via amicro valve (not shown). There are a plurality of cell storagemicrochannels 2006 connected between one surface of the mainmicrochannel 2004 and a waste microchannel 2008. Each of these cellstorage microchannels 2006 associated there with a filter 2010 disposedat the end thereof proximate to the waste microchannel 2008. Each of thecell storage microchannels 2006 has a size that will receive aparticular target cell having a particular dimension, such that thetarget cell will flow into the cell storage microchannel and cells ofsmaller size will pass through the associated filter 2010, which filter2010 is a microchannel with a diameter that is smaller than that of thetarget cell. This waste material will flow out through the waste gate ormicro valve (not shown) associated with the waste microchannel 2008. Bymaintaining a pressure differential between the main microchannel 2004and the waste microchannel 2008, the target cells will be stored withinthe cell storage channels 2006. Larger cells than the target cells inthe main microchannel 2004 will bypass the cell storage microchannels2006 and pass out of the waste gate associated with the mainmicrochannel 2004, keeping in mind that there is required to be a lowerpressure within the waste microchannel 2008 as compared to the mainmicrochannel 2004.

Referring now to FIG. 21, there is illustrated an embodiment of themicrofluidic chip utilizing micro valves as opposed to intermediatemicropumps. In this embodiment, there are illustrated a plurality ofinput wells 2102 for interfacing with an initial micropump 2104 to pumpfluid through a viewing window 2106 to a first reservoir 2108. Havingmultiple wells 2102 allows multiple samples to be input through theviewing window 2106 or to actually mix two different materials togetherfor flowing through the viewing window 2106 to the reservoir 2108. Thewaste gate 2110 can be provided at the reservoir connected thereto via awaste microchannel 2112 to allow air/bubbles to escape. A micropump 2114is operable to pump fluid from the reservoir 2108 to a main microchannelmanifold 2116. During this pumping operation, some type of pressurerelief is required which can either be provided via one of the pumps2104 being activated or a relief micro valve 2118 Interface with theinput end of the viewing window 2106 through a relief microchannel 2120.

Interfaced with the main microchannel manifold 2116 is a plurality ofdistribution micro valves 2124. These distribution micro valves 2124 canbe interfaced with various modules, as described above herein withrespect to FIGS. 17-20A/b. The only difference is that the associateddistribution pump 1624 has been replaced by a distribution valve 2124.Additionally, each of the parallel loaded testing reservoirs 312 can beindividually associated with one of the distribution valves 2124 toselectively certain ones thereof for testing. Since each one of thesetesting reservoirs 312, after selection, is required to be completelyfilled, by allowing individual selection of the testing reservoirs 312,certain ones thereof can be eliminated. It may be that, in pre-analyzingthe biofluid sample, it can be predetermined that certain ones of theassociated reagents in the reservoir 312 are not required thetesting/analysis step and can therefore be eliminated from the step offilling. This is opposed to the embodiment of FIG. 17, wherein all ofthe testing reservoirs 312 are complete the filled.

Referring now to FIGS. 22A-22B, there is illustrated cross-sectionalviews of a micro valve in an open and a closed position. The substrate402 has cover plate 440 disposed on top thereof. There are provided tomicrochannels 2202 and 2204 that are to be connected together with themicro valve. The microchannel 2202 is interfaced with a hole 2006 to thesurface of the cover plate 440 to an opening 2208. The microchannel 2204is interfaced to a hole 2210 to an opening 2212 in the cover plate 440.The micro valve has a fixed body 2214 with a membrane 2216 disposed onthe surface there above to define a pumping chamber 2218. The pumpingchamber 2218 has a hole 2220 interfacing the pumping chamber 2218 withthe opening 2208 on the cover plate 440. Similarly, the hole 2212 isinterfaced to the pumping chamber 2218 through a hole 2222. The membrane2216 is operable to reciprocate away from the surface of the fixed body2214 exposing the top of the hole 220 in the pumping chamber 2218 toallow fluid to flow through the pumping chamber 2218 and down throughthe opening 2222 through the cover plate 440 and through to themicrochannel 2204. In the closed position, the membrane 2216 is forceddown against the upper end of the hole 2220. A pneumatic cavity 2230 isdisposed above the membrane 2216 in a housing 2232 and interfaces with apneumatic source through a hose 2234. Thus, by drawing a vacuum in thepneumatic cavity 2230, the membrane 2216 will be pulled away from thehole 2220 to allow fluid to flow and, when pressurized air is forcedinto the pneumatic cavity 2230, and the membrane 2216 is forced down tothe surface of the fixed body 2214 to seal the opening 2224 in a closedposition.

Referring now to FIG. 23, there is illustrated a process flow forpreparing the biologic sample for the microfluidic chip 102. Thepreparation of the biologic sample can take many forms. In this example,the raw biologic sample can be preprocessed, depending upon the type ofsample that is being considered. For example, if blood is being tested,the Complete Blood Count (CBC) can be determined, as well as the WhiteBlood Cell Count (WBC), the liver functions and the kidney functions.For urinalysis, the sample can be prepared for testing for WBC's andnitrates, as well as proteins and Bilirubin. There are many well-knownprocesses for preparing biologic samples prior to testing. Once thebiologic sample has been prepared a, affinity labels are attachedthereto. Typically, there will be a vial 2302 provided with the biologicsample that is mixed with affinity labels in a vial 2304 resulting inthe vial 2304 containing a labeled sample. These labels are sometimesreferred to as “affinity labels” or “microspheres.” These functionalpolymeric microspheres typically have a diameter of less than 5 μm andhave been developed for use with immunological methods. The reagentswere initially used as visual markers to identify specific cell typesand analyze the distribution of cell surface antigens by scanningelectron microscopy. They have also been used, due to their inherentproperties, two separate labeled from unlabeled cells by techniques suchas centrifugation, a electrophoresis, magnetic chromatography andfluorescence cell sorting. The cells contained within the biologicsample are basically cells bearing defined antigens or receptors,ligands which bind with a high degree of selectivity an affinity tothese cell surface sites. The microspheres interact with the specificligand, which can allow for separation based upon the characteristicproperties of the microspheres. This allows for displaying of theselabeled cells with the target receptor or antigen with antibiotics orother ligands directly or indirectly bound to the microspheres. Specifictypes of microspheres or affinity labels can be the type that willfluoresce at a particular wavelength. Thus, specific cells can beidentified the optical techniques to identify target cells ordifferentiate between various types of, for example, bacterial cells andproteins, etc. This labeled sample is then disposed within the well 302on the microfluidic chip 102 for later processing.

Referring now to FIG. 24, there is illustrated a side cross-sectionalview of an RT-lamp. The RT-lamp is a Reverse Transcription Loop-mediatedisothermal Amplification device, which is an a technique for theamplification of RNA. This combines the advantages of the reversetranscription without of the LAMP technique. The LAMP technique is asingle to technique for the application of DNA. This technique is anisothermal nucleic acid application technique, in which a chain reactionis carried out at a constant temperature and does not require a thermalcycler. The target sequences animal five at a constant temperature usingeither two or three sets of primers and polymerase with high stranddisplacement activity in addition to a replication activity. Theaddition of the reverse transcription phase allows for the detection ofRNA and provides a one-step nucleic acid amplification method that isused to diagnose infectious diseases caused by bacteria or viruses.

FIG. 24 illustrates an example in which a multimode instrument 2401 iscoupled to a smartphone 2402. The smartphone 2402 includes an LED 2404and a camera 2406. The camera 2406 includes an image sensor, such as aCCD. The instrument 2400 includes a sample chamber 2408 for receiving anoptical assay medium. The optical assay medium comprises the labeledbiologic sample disposed within the viewing window 306 on themicrofluidic chip 102. The sample chamber 2408 may include a door 2410that prevents stray light from entering

The optical assay medium is positioned over a detection head 2412 in thesample chamber 2408. The instrument 2400 includes an optical output pathfor receiving an optical output from the optical assay medium in thesample chamber 2408 via the detection head 2412. The optical output pathmay include a multimode fiber 2414 that directs light from the detectionhead 2412 to a cylindrical lens 2416. The optical output path mayfurther include a wavelength-dispersive element, such as a diffractiongrating 2418, that is configured to disperse the optical output intospatially-separated wavelength components. The optical output path mayalso include other optical components, such as collimating lenses,filters, and polarizers.

The instrument 2401 can include a mount for removably mounting thesmartphone 2402 in a working position such that the camera 2406 isoptically coupled to the optical path, for example, in a predeterminedposition relative to the diffraction grating 2418. In this workingposition, the camera 2406 can receive at least a portion of thedispersed optical output such that different locations are received atdifferent locations on the image sensor.

The instrument 2401 may also include an input optical path for directinglight from a light source to the optical assay medium in the samplechamber 2408, for example, through the detection head 2412. In someinstances, the LED 2404 on the smartphone 2402 could be used as thelight source. To use the LED 2404 as the light source, the input opticalpath may include a collimating lens 2420 that receives light from theLED 2404 when the smartphone 2402 is mounted to the instrument 2400 inthe working position. The input optical path may further include amultimode fiber 2422 that directs the light from the collimating lens2420 to the detection head 2412. The input optical path may also includeother optical components, such as collimating lenses, filters, andpolarizers.

The instrument 2400 may also include an additional input optical paththat directs light form an internal light source, such as a laser 2424,to the optical assay medium in the sample chamber 2408. The additionalinput optical path may include a multimode optical fiber 2426, as wellas collimating lenses, filters, polarizers, or other optical components.

Referring now to FIG. 25, there is illustrated the view of the RT-lamp2401 with a microfluidics chip 102 disposed within the sample chamber2408.

Referring now to FIG. 26, there is illustrated a side view of the smartphone 2402 interfaced with the microfluidic chip 102 four imaging thesurface thereof, which is illustrated in a window view in FIG. 27. Thiswindow view illustrates the viewing window as a box 2702 in which theimage of the microfluidic chip 102 is displayed. The applicationautomatic the recognizes various markers 2704, 2706 and 2708 one threecorners thereof. This will allow orientation of the window with respectto the application. A box 2710 in phantom dashes will be oriented by theapplication running on the smart phone 2402. Once the box has beenoriented visually about the image of the microfluidic chip 102, thenprocessing can proceed. The processing is basically focusing upon thechip to gain the best optical image of the target sites. The targetsites are storage reservoirs 312 and 330, for example. Each of thesewill have a viewing well 318 associated there with an these viewingwells 318 will have, and one example, a process biologic sample havingaffinity labels associated there with that fluoresce. By recognizing theflorescence, the presence of the cell can be determined. The lack offlorescence indicates that the cell, a bacteria for example, has beendestroyed. This can be a positive test. By examining at each stage ofthe testing process the chip, a determination can be made as to resultsin essentially real time. This will be described in more detailhereinbelow. Once the image is believed to be in focus, then the usercan actually take the picture or the application cell can automaticallydetermine that the focus is adequate and take that. This is very similarto character recognition techniques that are utilized in recognizingfaces in camera images received by the phone.

Referring now to FIGS. 28A-28H, there are illustrated various images ofthe microfluidic chip 102 at different stages, this view being adiagrammatic view for simplicity. In this view, there is provided thesample well 2802 which then feeds into the viewing well 2804. Asdescribed hereinabove, there are multiple pumps that allow fluid to bemoved from the sample well 2802 over to the viewing well 2804 and theseare not shown force simplicity purposes. There is provided a multiplexer2802 which represents the micropumps/valves described hereinabove. Themultiplexer 2008 may be associated with one bank 2808 of reservoirs 2802for the parallel processing stage. These reservoirs 2012 correspond tothe reservoirs 312. This requires that the multiplexer 2806 distributefluid to a microchannel manifold 2810 and one testing phase. Themultiplexer 2006 also is connected via a plurality of microchannels to abank of reservoirs associated with the serial processing stage toselectively distribute fluid to one of the strings in a second testingphase. This bank of reservoirs includes the reservoirs 330 describedhereinabove. Each of these reservoirs 330 is arranged in series suchthat each has a valve or pump disposed there between. The multiplexer2806 also interfaces with a bank 2830 of reservoirs, these, in thisexample, associated with the serial testing/analysis stage and havingreservoirs 330 associated there with. In this example, there areprovided five test reservoirs in the bank 2808, wherein each of thesetest reservoirs has associated there with one serial string of testreservoirs 330 in the bank 2814 and one parallel loaded string ofreservoirs 330 in the bank 2820. Additionally, there is a separatetesting reservoir 2824 which could correspond to the cell storage areautilizing a chemostat described hereinabove, which is interfaced withmultiplexer 2806 through a microchannel 2826.

Referring now to FIGS. 28B-28H, there are illustrated various stages ofthe loading and analysis. FIG. 28B illustrates the first step in theprocess wherein the biologic sample is loaded into the viewing window2004. That the step in the process, the microfluidic chip is disposedwithin the RT lamp 2401 and analyzed to determine the number of cellsand the type of cells. If, for example, a certain bacteria were beingtested for on this particular microfluidic chip 102, the lack ofbacteria cells, as indicated by the particular affinity labels thatwould be attached to these particular bacteria cells, would indicatethat further testing is not required. However, if the correct cells arelabeled and the number of cells is at an appropriate level for testing,then the next step of the process is taken.

FIG. 28C illustrates a next step of the process wherein a portion of thecontents of the viewing well 2804 are transferred to all of thereservoirs 28 one two in the bank 2808, there being five reservoirs 2812disposed therein, the indirect dies that there could be more reservoirs2012 provided on the microfluidic chip 102. There will be a certainamount of time required for the pump associated with the multiplexer2808 to actually move the desire portion of the biologic sample throughthe manifold 2810 to the reservoirs 2812. As noted hereinabove, each ofthe reservoirs 2812 corresponds to the reservoirs 312, each having aserpentine microchannel 316 and a viewing reservoir 318 associated therewith. The micro pumps associated with the multiplexer 2806 and, nationwith the very small widths of the microchannels can require this processto take upwards of 10 or 20 minutes. After this period of time, themicrofluidic chip 102 can be imaged to determine if the cells have beendestroyed by the coating on the surfaces of the serpentine channel 316.(It should be noted that the viewing well 318 could also be coated). Ifthe cells are destroyed, this indicates that the reagent that coats thewalls of the microchannel associated with the reservoirs 312 reacted ina manner indicating self-destruction. However, any visual indication inthe viewing wells that can be a vehicle for discrimination betweeninteraction with the particular reagent coating the walls of theserpentine microchannels 316 will provide the ability for a decision tobe made as to which reagent is required for further testing.

FIG. 28D illustrates the next phase of operation, which is the phase inwhich the dosage level is term and. In the example above, the middlereservoir in the bank 2808 provided a trigger indication that triggereda decision to then test for dosage in the middle string within the bank2814. This will require a multiplexer 2806 two only transfer theremaining portion of the biologic sample from the viewing reservoir 2804into this particular string. As described hereinabove, this process willinvolve first passing of fluid to the first reservoir 330, which willtake a certain amount of time to actually pump the biologic samplethrough the microchannels into the viewing window 318. This can be amultiphase process, which requires viewing at each stage. In thisparticular example, the third stage of testing in this middle string inthe bank of reservoirs 2814 resulted in a perceivable result, i.e., alack of florescence, for example. At this point, the image will actuallyshow the perceivable result in both the bank 2808 and in the bank 2814.Thus, in the three phases of testing, the particular cells have been adefined, an indication has been provided as to which of multiplereagents that could possibly provide the desired therapeutic resultswould be the best choice for the patient and then the third phase oftesting provides the actual dosage of that determine reagent. It may bethat for ten individuals that had exactly the same symptoms andprocessed a similarly processed biologic sample for testing in the sameway with the microfluidic chip and the RT-lamp 2401 came up withdifferent results. Each individual's particular physiology can vary and,as such, the results could differ. In a typical medical environment, theparticular reagent of choice or drug of choice is determined by anindividual based upon various criteria. Since the medical professionaldoes not have the test directly in front of them, they might justprescribe, for example, a broad based antibiotic. They might follow thatup with testing of a biologic sample in a lab, which could take a numberof days just to determine exactly what bacteria is present and whatwould be the best antibiotic to use in order to attack this particularbacteria. Of course, the broad-spectrum antibiotic might have worked bythe time the test results come back. If not, these results might beuseful to the medical professional. However, these tests seldom if everactually focus in on the dosage that would be preferable for aparticular individual. If even the particular antibiotic could beidentified which was specific to that particular bacteria tested for andfound be present in the biologic sample, the dosage prescribed istypically a medium or high dosage, depending upon the criteria that themedical professional utilizes. However, the medical professionaltypically generalizes the physiology of any individual and maybe filtersthat based upon age, gender, etc. However, the individual physiology isnot taken into account.

With use of the present microfluidic chip 102, the entire testingprocess can be performed at the Point of Care (POC) in a relativelyshort amount of time. The result is not only the identification of thebest reagent to use but also the dosage. This is all accomplished with avery small amount of biologic sample.

FIG. 28E, there is illustrated a potential further processing that canbe provided. In this embodiment, the bank 2820 can have a differentmodification of the antibiotic that was determined from the testassociated with the bank 2808. This modification could be associatedwith the pH of the antibiotic, wherein it has been determined withrespect to some antibiotics that the pH of the antibiotic can affect theefficacy thereof. In this example, it can be seen that the thirdreservoir with respect to dosage is the one that is selected in the bank2814 but in the bank 2820, is the lowest dosage. Thus, the multiplexer2806 needs to first test the bank 2814 and then test the bank 2820.However, it should be understood that both the bank 2814 and the bank2020 could be identical, either serially loaded or parallel loaded, thecommonality being that they have a gradually increasing dose ofantibiotics that can be tested for.

FIGS. 28F-28G, there are illustrated two additional examples of twodifferent patients with substantially the same symptoms and utilizingsubstantially the same process for preparing the biologic sample. Withrespect to FIG. 28F, the fifth reservoir and the antibiotic associatedthere with exhibited the highest efficacy at the highest dose as todestroying the particular bacteria, in the example of the bacteria. Theassociated dosage determined from testing the biologic sample in thebank 2814 was considered to be the second level of dosage. In the bank2020, the third level of dosage was considered to be the lowest dose.With respect to FIG. 28G, the first reservoir and the antibioticassociated there with was considered to have the highest efficacy withrespect to dealing with the particular bacteria and it was the lowestdose in that case when tested in the bank 2814, as compared to thefourth level dosage in the bank 2820. It can be seen thus that differentpatients will have different “fingerprints” associated with the testingof the same biologic sample repaired and substantially same way.

FIG. 28H, there is illustrated an alternate embodiment wherein the testperformed at the bank 2808 resulted in a slight ambiguity in that thebacteria were killed in two other reservoirs. In this case, theindication would be that either of these antibiotics would work againstthis particular strain of bacteria. Thus, the next phase the test wouldrequire the multiplexer 2808 to distribute the contents of the reservoir2804 through the microchannels to actually two different strings. Thus,for this type of test to be carried out, it is important that there besufficient volume in the viewing window 2804, i.e., sufficient amount ofbiofluid introduced to the well 2802, in order to fill both of thesereservoirs and allow the testing to progress down to the highest dosagelevel in either or both of the banks 2814 and 2820. The results of thistest show that, for the rightmost reservoir in the bank 2808 having beendetermined to be effective at the highest dose, the next of the lastdosage was required in order to achieve the desired results, whereas thenext to the left reservoir in the bank 2808 having been determined to beeffective at the highest dose required only the smallest dose to achievethe results. Therefore, this test shows that, although two antibioticswould work, one would actually work with the lower dose.

It should also be understood that, in addition to the test beingdifferent for the same strain of bacteria in a biologic prepared mensubstantially the same way, it should also be understood that thisparticular set of results could be different for different strains ofthe same bacteria. It may be that, for one strain, one antibiotic wouldwork at a particular dose and, for another strain of the same bacteria,a different antibiotic work or just a different dose of the sameantibiotic. The microfluidic chip described and disclosed in the presentdisclosure allows this determination to be made utilizing a singlesample in a parallel/serial testing method at the POC wherein the firststep or phase of selection is made among a plurality of potentialantibiotics that could arguably target different bacteria and, once adetermination is made at the first phase, then the next and serialdecision is made to determine dosage at a second phase.

Referring now to FIG. 29, there is illustrated a flowchart depicting theoverall analysis process. The process is initiated at a block 2902 andthen proceeds to a block 2904 wherein the biologic sample is prepared.As described hereinabove, this preparation involves labeling the cellswithin the biologic sample so that they can be discriminated between oridentified. It may be that there are a number of different types ofcells such as bacteria of different strains and types, proteins, etc.Different affinity labels can be applied such that multiple cells ofdifferent types can be identified. The process then flows to a block2906 wherein the biologic sample is placed into the sample well and thenpassed on to the viewing well. At this point, the microfluidic chip isplaced into the RT-lamp and optically analyzed, as indicated by processblock 2908. It is at this point in the testing phase that theidentification process will identify the potential target cells. Sinceeach of the microfluidic chips has a finite number of reservoirsassociated there with for the purpose of testing, the coating is appliedto these particular reservoirs for the specific antibiotics or reagentsto be tested may not be useful for testing the particular cellularstructures that have been identified at this step in the process.However, it should be understood that the number of different banks oftesting reservoirs that can be provided on a particular microfluidicchip can be expandable and the could actually be provided for multipledifferent types of reagents. For example, one set of testing banks maybe associated with UTI and another associated with streptococcalbacteria. Recognizing these at this step in utilizing them with amicrofluidic chip that can test for both types of bacteria will allowthe particular biologic sample, which is quite small, to be routed tothe appropriate reservoirs for testing for that specific identifybacteria.

The decision to proceed is determined at a decision block 2910 and, iftesting can proceed with the current microfluidic chip, the processproceeds to a block 2911 to select the particular test that are to beperformed. The process then proceeds to sequence through the tests, asindicated by a block 2914. This sequencing sequences through the variousphases, with the initial test being selected first, as indicated byblock 2916. In the above examples, this is the first parallel phase todetermine which among several reagents is most effective against theparticular cellular structure of interest. The process and proceeds to ablock 29 eight teen in order to analyze the results of this initial testand then to a decision block 2920 to determine if more tests arerequired or if this is the only test. If the test is negative at thisstage and none of the reagents provides any effectiveness indication,the process is terminated or, if this is the last test, the process isterminated. The process, if continued, then selects the next test in thesequence and proceeds back to the input of the block 2918 to continuesequencing through the tests.

Referring now to FIG. 30 come there is illustrated a flowchart for thetesting process. This is initiated at a block 3002 and then proceeds toa block 3004 two first pump a portion of the biologic sample stored inthe viewing window through to the parallel reservoirs and load all ofthe parallel reservoirs for testing/analysis. This may take upwards of10 or 20 minutes, due to the fact that the micropumps utilized arerelatively slow and the diameter of the microchannels is small, thusrestricting high flow rates. The process then flows to decision block3006 to determine if there is been any positive result, i.e., is thereany indication that any of the reagents provide an effectivenessindication, either through some color change or the lack of colorindicating the destruction of the cells. If there is no result, then theprocess is terminated in the process flows to a function block 3008 twoselect the next test path that is associated with the antibiotic havingbeen tested as being effective in the first phase of operation/testing.A process block 3010 and indicates that a graded dosage test path isselected, either the one for loading parallel or the one or loadingserially. It should be understood that the parallel loaded graded dosagetest path requires all of the reservoirs to be completely filled fromthe reservoir associated with the viewing window. The serial path, bycomparison, allows all of the contents of the viewing window in thereservoir associated there with to be disposed in each reservoir andthen sequentially transferred to the next reservoir down the chain andat the higher dosage. However, it should be understood that the systemcan be configured such that the first reservoir at the lowest dosage isloaded with only a portion of the contents of the viewing window and thereservoir associated there with, analyzed and then a micro valve gateopened to allow the micropumps for pumping fluid to the serial path tooperate to continue pushing more biofluid through the first reservoir,thus filling the second and reservoir and so on. In this process,sufficient biofluid must be contained within the viewing window and thereservoir associated there with in order to allow for filling of all ofthe reservoirs down to the highest dosage rate associated with thatserial string.

In the process, the serial string will first select the lowest gradeddose reservoir and a process block 3012 and then pump biofluid to thefirst reservoir and a process block 3014, analyze the results a processblock 3016, understanding that it could take 10 to 20 minutes to filleach reservoir. A determination is made at a decision block 3018 as towhether there is a positive result, i.e., was there and an effectivenessdetermination made at this point, and, if so proceed to a decision block3020 to determine if there are any higher concentrations to be testedfor. If so, the next reservoir selected by opening gate or activating amicropump, as indicated by a process block 3022, and the proceed back tothe process block 3014 in order to pump to this reservoir.

In the parallel process, a process block 3026 indicates an operationwherein the micropump pumps sufficient biofluid material to the parallelrated reservoirs to fill all of the reservoirs and into a process block3028 in order to analyze the results.

Referring now to FIG. 31 there is illustrated a perspective view of amodular microfluidic chip system 3102. The system 3102 includes amicrofluidic distributor chip 3104. This chip is similar to themicrofluidic chip 102 described hereinabove, with the exception thatonly a portion of the microchannels and wells are contained thereon. Inthis embodiment, the sample well 302, the first micropump 304 and theviewing well 306 are all contained on the surface of the distributorchip 3104. The output side of the viewing well 306 is interfaced with amicro pump distributor 3108. In this embodiment, there are provided onone side of the distributor chip 3104 three ports 3110, 3112 and 3116,each interfaced with the micropump distributor 3108 through a separatemicrochannel. Certainly, on the other side of the distributor chip 3104are provided three complementary ports 3116, 3118 and 3120, eachinterfaced with the micro-pump distributor 3108. At one end ofdistributor chip 3104 are provided two ports 3122 and 3124, eachinterfaced with the micro-pump distributor 3108. The three ports3110-3114 all disposed on the edge of the distributor chip 3104 andinterfaced with a slot 3126. Similarly, the three ports 3116-3120 aredisposed on the outside of the chip and interfaced with a slot 3128.Similarly, the portion 3122 and 3124 are disposed on the edge of thedistributor chip 3104 and interfaced with a slot 3130.

Each of the slots 3126-3130 is keyed to receive a module. There isillustrated a single module 3132. The module 3132 is similar to themicrofluidic chip 102 and that it has microfluidic channels, testingreservoirs, etc. disposed thereon. Any combination of testingreservoirs, micropumps, valves, etc. can be disposed on this module3132. At the edge of this module are provided three semicircular keyedsections 3134, 3136 and 3138. Each of the keyed sections 3134-3138 isassociated with a respective one of the ports 3116-3120 andcorrespondingly shaped portions of the slot 3128. There arecomplementary ports 3138, 3140 and 3142 in the respective keyed sections3138-3134 that correspond to and interface with the ports 3116-3120,respectively. When the module 3132 is fully inserted within the slot3128, the ports 3138-3142 have a fluid interface with the ports3116-3120. Thus, modules can be interchanged to provide differentfunctions. Additionally, if one module were associated with, forexample, the parallel loading operation wherein multiple differentreagents at the high concentration level and associated with oneparticular type of bacteria were contained within different associatedreservoirs, another module with a similar structure could have differentreagents contained therein that are associated with a another type ofbacteria. Thus, in the first phase of testing, utilizing the RT-lamp,the type of bacteria could be determined and the testing/analysis stepperformed the next phase of testing could then be determined so as toenable the correct module to be selected.

Referring now to FIG. 32, there is illustrated a cross-sectional view ofthe interface between the ports on the distributor chip 3104 and on themodule 3132. The distributor to 3104 contains a base substrate 3202 uponwhich is disposed in the upper substrate 3204 four containingmicrochannels. Disposed above the substrate 3204 is a cover plate 3206.Illustrated within the substrate he 204 is a single microchannel 3208which extends along the surface of the substrate 3204 and then extendsdownward through a vertical microchannel 3210 two an opening or port3212. The opening is disposed on the upper surface of the slot 3128,this being a surface 3150. The slot 3128 has a corresponding bottomsurface 3152 to provide an opening for receiving the module 3132. Themodule 3132 has a substrate 3154 containing microchannels, a singlemicrochannel 3156 illustrated. The module 3132 has a cover plate 3158disposed above the substrate 3154 with a vertical microchannel 3160disposed there through to an opening 3162. This opening 3162 is operableto interface with the opening 3212 when the module 3132 is inserted inthe slot 3128. Some type of flexible membrane 3166 could be provided onthe cover plate 3158 to provide a fluid seal. However, a tight fitbetween the cover plate 3158 and the bottom surface of the substrate3204 should provide an adequate seal for the microfluidic channel formedbetween the two devices.

Referring now to FIG. 33, there is illustrated a system level top viewof the modular system. There are illustrated multiple modules 3132having three ports associated there with for interfacing with either ofthe slots 3128 or 3126. There is provided a single two port module 3170for interfacing with the slot 3130. These modules can each havedifferent functionality associated there with and are operable to beinserted into the slots to very the functionality of the overall chip.

Referring now to FIG. 34, there is illustrated top view of thedistributor chip 3104 illustrating the micropump distributor 3108illustrating that a plurality of micropumps 3174 are disposed aroundeach of the edges to interface with the various ports 3110-3124. Thereis provided a single pump for pumping from the well 306 into a centraldistributor manifold 3176.

It will be appreciated by those skilled in the art having the benefit ofthis disclosure of a Modular Parallel/Serial Dual Microfluidic Chip. Itshould be understood that the drawings and detailed description hereinare to be regarded in an illustrative rather than a restrictive manner,and are not intended to be limiting to the particular forms and examplesdisclosed. On the contrary, included are any further modifications,changes, rearrangements, substitutions, alternatives, design choices,and embodiments apparent to those of ordinary skill in the art, withoutdeparting from the spirit and scope hereof, as defined by the followingclaims. Thus, it is intended that the following claims be interpreted toembrace all such further modifications, changes, rearrangements,substitutions, alternatives, design choices, and embodiments.

What is claimed is:
 1. A modular microfluidic chip system for testing atreatment agent for a predetermined biologic material, comprising: adistributor module including: an input for receiving a biologic sample,the biologic sample containing the predetermined biologic material thatmust be treated via one of a plurality of treatment agents; a firstreservoir for holding the biologic sample containing the predeterminedbiologic material; a plurality of micro-pumps for pumping a portion ofthe biologic sample from the first reservoir into a connected module; afirst module interconnected with the first module and including a firstplurality of parallel pathways each for testing a treatment agent of theplurality of treatment agents and determining a treatment efficacy forthe predetermined biologic material within the biologic sample withrespect to the treatment agent; wherein a first micro-pump of theplurality of micro-pumps pumps a first portion of the biologic sampleinto each of the first plurality of parallel pathways within the firstmodule from the first reservoir; a first module connector for removeablyconnecting the first module to the modular fluidic chip system, whereinthe plurality of treatment agents tested by the system may be selectedby selecting the first module associated with the first moduleconnector; a second module including a second plurality of parallelpathways each for determining a dosage level of a particular one of theplurality of treatment agents with respect to the predetermined biologicmaterial; wherein a second plurality of micro-pumps of the plurality ofmicro-pumps pumps a second portion of the biologic sample into aselected one of the second plurality of pathways responsive to a controlinput indicating the treatment agent providing the best treatmentefficacy of the predetermined biologic material; and a second moduleconnector for removeably connecting the second module to the distributormodule, wherein treatment agent and the plurality of dosage levelstested by the system may be selected by selecting the second moduleassociated with the second module connector.
 2. The system of claim 1,wherein the first plurality of parallel pathways further comprises: aplurality of second reservoirs for holding the portion of the biologicsample treated with one of a plurality of treatment agents; and aplurality of micro-channels, each of the plurality of micro-channelsinterconnecting the first reservoir to the plurality of secondreservoirs, each of the plurality of micro-channels having a portionthereof interiorly coated with one of the plurality of treatment agentsfor applying the treatment agent to the predetermined biologic materialpassing through the portion of the micro-channel.
 3. The system of claim2, wherein each of the plurality of micro-channels include a serpentineportion for the portion of the micro-channel.
 4. The system of claim 2,wherein the first micro-pump further pumps the portion of the biologicsample through the plurality of micro-channels into the plurality ofsecond reservoirs.
 5. The system of claim 2, further including a secondplurality of reading windows each associated with one of the pluralityof second reservoirs for enabling a view of effects caused byapplication of the treatment agent to the biologic sample.
 6. The systemof claim 1, wherein each of the second plurality of pathways furthercomprises a plurality of testing modules each for applying a differentdosage level of one of the plurality of treatment agents to the biologicmaterial within the biologic sample.
 7. The system of claim 6, whereineach of the plurality of testing modules further comprises: a thirdreservoir for holding the second portion of the biologic sample treatedwith one of a plurality of treatment agents; and a micro-channelinterconnecting the first reservoir to the third reservoir, themicro-channel including a portion interiorly coated with one of theplurality of treatment agents for applying the treatment agent to thepredetermined biologic material passing through the micro-channel at thedifferent dosage level.
 8. The system of claim 7, wherein the pluralityof testing modules are connected in series to test an efficacy of aplurality of dosage levels of the testing agent one at a time.
 9. Thesystem of claim 7, wherein the plurality of testing modules areconnected in parallel to test an efficacy of a plurality of dosagelevels of the testing agent at a same time.
 10. The system of claim 1further comprising a cell counter associated with the first readingwindow for applying an affinity label to cells of the detected biologicmaterial within the biologic sample.
 11. A microfluidic chip system fortesting a treatment agent for a predetermined biologic material,comprising: a distributor module including: an input for receiving abiologic sample, the biologic sample containing the predeterminedbiologic material that must be treated via one of a plurality oftreatment agents; a first reservoir for holding the biologic samplecontaining the predetermined biologic material; a plurality ofmicro-pumps for pumping a portion of the biologic sample from the firstreservoir into a connected module; a first module including a firstplurality of parallel pathways each for testing a treatment agent of theplurality of treatment agents and determining a treatment efficacy forthe predetermined biologic material within the biologic sample withrespect to the treatment agent, the first plurality of parallel pathwaysfurther comprising: a plurality of second reservoirs for holding aportion of the biologic sample treated with one of a plurality oftreatment agents; a plurality of micro-channels, each of the pluralityof micro-channels interconnecting the first reservoir to the pluralityof second reservoirs, each of the plurality of micro-channels having aportion thereof interiorly coated with one of a plurality of treatmentagents for applying the treatment agent to the predetermined biologicmaterial passing through the portion of the micro-channel; a firstmodule connector for removeably connecting the first module to thedistributor module, wherein the plurality of treatment agents tested bythe system may be selected by selecting the first module associated withthe first module connector; wherein a first micro-pump of the pluralityof micro-pumps pumps the portion of the biologic sample into each of thefirst plurality of parallel pathways from the first reservoir; a secondmodule including a second plurality of parallel pathways each fordetermining a dosage level of a particular one of the plurality oftreatment agents with respect to the predetermined biologic material,wherein the second plurality of parallel pathways further comprises: aplurality of testing modules each for applying a different dosage levelof one of the plurality of treatment agents to the biologic materialwithin the biologic sample, wherein each of the plurality of testingmodules further comprises: a third reservoir for holding the secondportion of the biologic sample treated with one of a plurality oftreatment agents at a selected dosage level; a micro-channelinterconnecting the first reservoir to the third reservoir, themicro-channel including a portion interiorly coated with one of theplurality of treatment agents for applying the treatment agent at thedifferent dosage level to the predetermined biologic material passingthrough the micro-channel; a second module connector for removeablyconnecting the second module to the distributor module, whereintreatment agent and the plurality of dosage levels tested by the systemmay be selected by selecting the second module connected to the secondmodule connector; and wherein a second micro-pump of the plurality ofmicro-pumps pumps the second portion of the biologic sample into theselected one of the micro-channels responsive to a control inputindicating the treatment agent providing the best treatment efficacy ofthe predetermined biologic material.
 12. The system of claim 11, whereineach of the plurality of micro-channels include a serpentine portion forthe portion of the micro-channel.
 13. The system of claim 11, whereinthe first micro-pump further pumps the portion of the biologic samplethrough the plurality of micro-channels into the plurality of secondreservoirs.
 14. The system of claim 11, further including a secondplurality of reading windows each associated with one of the pluralityof second reservoirs for enabling a view of effects caused byapplication of the treatment agent to the biologic sample.
 15. Thesystem of claim 11, wherein the plurality of testing modules areconnected in series to test an efficacy of a plurality of dosage levelsof the testing agent one at a time.
 16. The system of claim 11, whereinthe plurality of testing modules are connected in parallel to test anefficacy of a plurality of dosage levels of the testing agent at a sametime.
 17. The system of claim 11 further comprising a cell counterassociated with the first reading window for applying an affinity labelto cells of the detected biologic material within the biologic sample.18. A system for testing a treatment agent for a predetermined biologicmaterial, comprising: a distributor module including: an input forreceiving a biologic sample, the biologic sample containing a uniquecombination of a predetermined biologic material that must be treatedvia one of a plurality of treatment agents and a patient's biologicmaterial; a first reservoir for holding the biologic sample containingthe unique combination of the predetermined biologic material; aplurality of micro-pumps for pumping a portion of the biologic samplefrom the first reservoir into a connected module; a first moduleincluding a first plurality of testing pathways for testing a firstportion of the biologic sample containing the unique combination of thepredetermined biologic material and the patient's biologic material,each of the plurality of first parallel testing pathways applying adifferent treatment agent of a plurality of treatment agents to thefirst portion of the biologic sample; a first module connector forremoveably connecting the first module to the distributor module,wherein the plurality of treatment agents tested by the system may beselected by selecting the first module associated with the first moduleconnector; a second module including a second plurality of testingpathways for testing a second portion of the biologic sample containingthe unique combination of the predetermined biologic material and thepatient's biologic material in a selected pathway of the secondplurality of testing pathways, the selected pathway selected responsiveto a control input responsive to results of the first plurality oftesting pathways, the selected pathway applying at least one dosagelevel of a treatment agent to the second portion of the biologic sample;and a second module connector for removeably connecting the secondmodule to the distributor module, wherein treatment agent and theplurality of dosage levels tested by the system may be selected byselecting the second module associated with the second module connector.19. The system of claim 18, wherein the second plurality of treatmentpathways further applies the treatment agent in series at the pluralityof different dosage levels to test an efficacy of the plurality ofdosage levels one at a time.
 20. The system of claim 18, wherein thesecond plurality of treatment pathways further applies the determinedtreatment agent at the plurality of dosage levels in parallel to test anefficacy of the plurality of dosage levels at a same time.
 21. A methodfor testing a treatment agent for a predetermined biologic materialusing a modular testing system, comprising: connecting a first moduleproviding testing of the plurality of treatment agents to a distributormodule; connecting a second module providing testing of the plurality oftreatment agents at a plurality of predetermined dosage levels to adistributor module; receiving at the distributor module a biologicsample, the biologic sample containing the predetermined biologicmaterial that must be treated via one of a plurality of treatmentagents; holding the biologic sample containing the predeterminedbiologic material within a first reservoir in the distributor module;pumping a portion of the biologic sample from the first reservoir in thedistributor module into each of a first plurality of parallel pathwaysin the first module; applying within the first module a treatment agentof a plurality of treatment agents within each of the first plurality ofparallel pathways to the portion of the biologic sample within theparallel pathway; pumping a second portion of the biologic sample fromthe first reservoir in the distributor module into a selected secondparallel pathway associated with the selected treatment agent of asecond plurality of parallel pathways within the second module using asecond micro-pump, the selected second pathway responsive to a controlinput; and applying within the second module the selected treatmentagent at a plurality of different dosage levels within the selectedsecond parallel pathway to the second portion of the biologic samplewithin the second parallel pathway.
 22. The method of claim 21, whereinthe step of applying a treatment agent further comprises pumping thebiologic sample through a plurality of micro-channels interconnectingthe first reservoir with a plurality of second reservoirs to apply theplurality of treatment agents, wherein one of the plurality of treatmentagents are applied in each of the plurality of micro-channels.
 23. Themethod of claim 21, wherein the step of applying the selected treatmentagent further comprises pumping the second portion of the biologicsample through a second micro-channels to a second reservoir to applythe selected of treatment agent at one of a plurality of dosage levelsto the second portion of the biologic sample.
 24. The method of claim21, wherein the step of applying the selected treatment agent furthercomprises applying the selected treatment agent in series at theplurality of different dosage levels to test an efficacy of theplurality of dosage levels one at a time.
 25. The method of claim 21,wherein the step of applying the selected treatment agent furthercomprises applying the selected treatment agent at the plurality ofdifferent dosage levels in parallel to test an efficacy of the pluralityof dosage levels at a same time.
 26. The method of claim 21 furthercomprising the step of applying an affinity label to cells of thedetected biologic material within the biologic sample using a cellcounter.